EphA4/ephrinA3 reverse signaling induced Müller cell gliosis and production of pro-inflammatory cytokines in experimental glaucoma

Lin-Jie Xu; Hong-Ning Wang; Han Zhou; Shu-Ying Li; Fang Li; Yanying Miao; Bo Lei; Xing-Huai Sun; Feng Gao; Zhongfeng Wang

doi:10.1016/j.brainres.2022.148204

EphA4/ephrinA3 reverse signaling induced Müller cell gliosis and production of pro-inflammatory cytokines in experimental glaucoma

Brain Res. 2023 Feb 15:1801:148204. doi: 10.1016/j.brainres.2022.148204. Epub 2022 Dec 15.

Authors

Lin-Jie Xu¹, Hong-Ning Wang², Han Zhou³, Shu-Ying Li⁴, Fang Li⁵, Yanying Miao⁶, Bo Lei⁷, Xing-Huai Sun⁸, Feng Gao⁹, Zhongfeng Wang¹⁰

Affiliations

¹ State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: 16111520007@fudan.edu.cn.
² State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: 18111520008@fudan.edu.cn.
³ State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: 19111520010@fudan.edu.cn.
⁴ State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: 20111520020@fudan.edu.cn.
⁵ State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: fanglee@fudan.edu.cn.
⁶ State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: yymiao@fudan.edu.cn.
⁷ Institute of Neuroscience and Third Affiliated Hospital, Henan Provincial People's Hospital, Henan Eye Institute, Henan Eye Hospital, People's Hospital of Zhengzhou University, Zhengzhou University, Zhengzhou 450003, China.
⁸ Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, NHC Key Laboratory of Myopia, Shanghai Key Laboratory of Visual Impairment and Restoration, Fudan University, Shanghai 200031, China. Electronic address: xhsun@shmu.edu.cn.
⁹ Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, NHC Key Laboratory of Myopia, Shanghai Key Laboratory of Visual Impairment and Restoration, Fudan University, Shanghai 200031, China. Electronic address: fenggao@fudan.edu.cn.
¹⁰ State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai 200032, China. Electronic address: zfwang@fudan.edu.cn.

PMID: 36529265
DOI: 10.1016/j.brainres.2022.148204

Abstract

Previous work showed that ephrinA3/EphA4 forward signaling contributed to retinal ganglion cell (RGC) damage in experimental glaucoma. Since up-regulated patterns of ephrinA3 and EphA4 were observed in Müller cells and RGCs, an EphA4/ephrinA3 reverse signaling may exist in Müller cells of chronic ocular hypertension (COH) retina. We investigated effects of EphA4/ephrinA3 reverse signaling activation on Müller cells in COH retina. Intravitreal injection of the ephrinA3 agonist EphA4-Fc increased glial fibrillary acidic protein (GFAP) levels in normal retinas, suggestive of Müller cell gliosis, which was confirmed in purified cultured Müller cells treated with EphA4-Fc. These effects were mediated by intracellular STAT3 signaling pathway as phosphorylated STAT3 (p-STAT3) levels and ratios of p-STAT3/STAT3 were significantly increased in both COH retinas and EphA4-Fc intravitreally injected retinas, as well as in EphA4-Fc treated purified cultured Müller cells. The increase of GFAP protein levels in EphA4-Fc-injected retinas and EphA4-Fc treated purified cultured Müller cells could be partially eliminated by stattic, a selective STAT3 blocker. Co-immunoprecipitation results testified to the presence of interaction between ephrinA3 and STAT3/p-STAT3. In addition, intravitreal injection of EphA4-Fc or EphA4-Fc treatment of cultured Müller cells significantly up-regulated mRNA and protein contents of pro-inflammatory cytokines. Moreover, intravitreal injection of EphA4-Fc increased the number of apoptotic RGCs, which could be reversed by the tyrosine kinase blocker PP2. Overall, EphA4/ephrinA3 reverse signaling may induce Müller cell gliosis and increases release of pro-inflammatory factors, which could contribute to RGC death in glaucoma. Inhibition of EphA4/ephrinA3 signaling may provide an effective neuroprotection in glaucoma.

Keywords: Chronic ocular hypertension; EphA4/ephrinA3 reverse signaling; GFAP; Glaucoma; Müller cells; Pro-inflammatory factors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cytokines / metabolism
Ependymoglial Cells* / metabolism
Ephrin-A3 / metabolism
Glaucoma*
Gliosis / metabolism
Humans
Receptor, EphA4 / metabolism
Signal Transduction / physiology

Substances

Cytokines
Ephrin-A3
Receptor, EphA4